Control of viral-host membrane fusion with hydrogen bond surrogate-based artificial helices

a technology of surrogate-based artificial helices and viralhosts, which is applied in the direction of drug compositions, peptide sources, peptide/protein ingredients, etc., can solve the problem that not all patients are responsive to existing therapies, and achieve the effect of inhibiting viral infectivity

Active Publication Date: 2014-10-28
NEW YORK UNIV
View PDF35 Cites 29 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, not all patients are responsive to existing therapies, and the virus develops resistance to most, if not all, known agents.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Control of viral-host membrane fusion with hydrogen bond surrogate-based artificial helices
  • Control of viral-host membrane fusion with hydrogen bond surrogate-based artificial helices
  • Control of viral-host membrane fusion with hydrogen bond surrogate-based artificial helices

Examples

Experimental program
Comparison scheme
Effect test

example 1

Synthesis of Peptides 1-11 and Characterization of Peptides 1-12

[0101]Peptides 1-11 were synthesized as shown in Scheme 1. Peptides 1 (i.e., AcMTWMEWDREINNYT-NH2 (SEQ ID NO: 14)), 10 (i.e., AcMTWEEWDKKIEEYTKKI—NH2 (SEQ ID NO: 15)), and 11 (i.e., peptide IZN17, AcIKKEIEAIKKEQEAIKKKIEAIEKLLQLTVWGIKQLQARIL-NH2 (SEQ ID NO: 16)), and resin-bound bis-olefins (17) were synthesized by conventional Fmoc solid phase chemistry on Rink amide HMBA resin (NovaBiochem), 0.05-0.15 mmol scale, with appropriate substitutions of N(allyl)-dipeptides 13-16 and 4-pentenoic acid. In each coupling step, the Fmoc group was removed by treatment with 20% piperidine in NMP (2×20 min). The next Fmoc amino acid (4 equiv.) in the sequence was activated with HBTU (3.6 equiv.) in a 5% DIPEA / NMP solution for 15 minutes, added to the resin bearing the free amine. The resulting mixture was shaken for 60 minutes. The coupling efficiency for each step was monitored by ninhydrin test. After the peptide was assembled on t...

example 2

Circular Dichroism Spectroscopy of Peptides 1-10

[0106]CD spectra were recorded on an AVIV 202SF CD spectrometer equipped with a temperature controller using 1 mm length cells and a scan speed of 5 nm / min. The spectra were averaged over 10 scans with the baseline subtracted from analogous conditions as that for the samples. The samples were prepared in 0.1× phosphate buffered saline (13.7 mM NaCl, 1 mM phosphate, 0.27 mM KCl, pH 7.4), containing 10% trifluoroethanol, with the final peptide concentration of 50-100 μM. The concentrations of unfolded peptides were determined by the UV absorption of the tyrosine residue at 276 nm in 6.0 M guanidinium hydrochloride aqueous solution. The helix content of each peptide was determined from the mean residue CD at 222 nm, [θ]222 (deg cm2 dmol−1) corrected for the number of amino acids. Percent helicity was calculated from the ratio [θ]222 / [θ]max, where [θ]max=(−44000+250T)(1−k / n), with k=4.0 and n=number of residues. For details on θmax calcula...

example 3

Affinity of Peptide 12 for IZN17

[0107]The relative affinity of peptides 1-10 for IZN17 (i.e., peptide 11) was determined using a fluorescence polarization-based competitive binding assay (Eckert & Kim, “Design of Potent Inhibitors of HIV-1 Entry from the gp41 N-Peptide Region,”Proc. Nat'l Acad. Sci. U.S.A. 98:11187-92 (2001); Stephens et al., “Inhibiting HIV Fusion with a β-Peptide Foldamer,”J. Am. Chem. Soc. 127:13126-7 (2005), which are hereby incorporated by reference in their entirety) with fluorescein-labeled peptide 12, shown in FIG. 13. FIG. 14 is a graph showing the saturation binding curve of fluorescein-labeled peptide 12 with IZN17 in PBS buffer at 25° C.

[0108]All samples were prepared in 96-well plates in 1× phosphate buffered saline (137 mM NaCl, 10 mM phosphate, 2.7 mM KCl, pH 7.4) with 0.1% pluronic F-68 (Sigma). A solution of 25 μM IZN17 and 15 nM fluorescein-labeled peptide 12 was incubated at 25° C. After 1 hour, appropriate concentrations (10 nM to 500 μM) of the ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
temperatureaaaaaaaaaa
temperatureaaaaaaaaaa
temperatureaaaaaaaaaa
Login to view more

Abstract

The present invention relates to a peptide having one or more stable, internally-constrained HBS α-helices, where the peptide mimics at least a portion of a class I C-peptide helix or at least a portion of a class I N-peptide helix of a viral (e.g., HIV-I) coiled-coil assembly. Methods of inhibiting viral infectivity of a subject by administering these peptides are also disclosed.

Description

CROSS-REFERENCE[0001]This application claims the benefit of U.S. Provisional Application No. 61 / 018,118, filed Dec. 31, 2007, which application is incorporated herein by reference.STATEMENT AS TO FEDERALLY SPONSORED RESEARCH[0002]This invention was made with government support under grant numbers GM073943 and AI42382, both awarded by the National Institutes of Health. The government has certain rights in this invention.BACKGROUND OF THE INVENTION[0003]Enveloped viruses depend upon fusion between the viral membrane and a host cell membrane (the plasma membrane or an intracellular membrane, depending upon the specific virus) for delivery of viral genetic material to the host cell, thereby initiating infection of the host cell (Kielian & Ray, “Virus Membrane-fusion Proteins: More Than One Way to Make a Hairpin,”Nat. Rev. 4:67-76 (2006), which is hereby incorporated by reference in its entirety). This membrane-fusion reaction relies on virus membrane-fusion proteins (Kielian & Ray, “Vir...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Patents(United States)
IPC IPC(8): C07K14/05C07K7/08C07K14/005A61K38/00
CPCC12N2740/16122C07K14/005A61K38/00A61P31/12A61P31/14A61P31/16A61P31/18A61P31/20A61P31/22
Inventor WANG, DEYUNARORA, PARAMJIT
Owner NEW YORK UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap